Equine Infectious Anemia Viral Vector-Mediated Codelivery of Endostatin and Angiostatin Driven by Retinal Pigmented Epithelium-Specific VMD2 Promoter Inhibits Choroidal Neovascularization
Equine Infectious Anemia Viral Vector-Mediated Codelivery of Endostatin and Angiostatin Driven by Retinal Pigmented Epithelium-Specific VMD2 Promoter Inhibits Choroidal Neovascularization
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DOI：
10.1089/hum.2008.046
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发表时间：
2009-01-01
影响因子：
4.2
通讯作者：
Campochiaro, Peter A.
中科院分区：
文献类型：
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作者：
Kachi, Shu;Binley, Katie;Campochiaro, Peter A.
马传染性贫血病毒（EIAV）是一种不会引起人类疾病的非灵长类慢病毒。在小鼠视网膜下注射重组EIAV慢病毒载体，其中lacZ由CMV启动子驱动（EIAV CMV lacZ），导致lacZ在视网膜色素上皮（RPE）细胞和其他一些细胞（包括神经节细胞）中快速和强烈表达，导致视神经内存在5-溴-4-氯-3-吲哚- β -d-半乳糖苷。将卵黄样黄斑营养不良（VMD2）基因中的RPE特异性启动子替换为CMV启动子导致LacZ的表达延长（至少1年），该表达仅限于RPE细胞，尽管与CMV启动子相比减少了6- 10倍。同样，在注射了EIAV VMD2 Endo（FLAG）载体的眼睛中检测到的FLAG标记的内皮抑制素的数量与注射了表达内皮抑制素和血管抑制素的载体的眼睛中检测到的数量相似[EIAV VMD2 Endo(FLAG)/Angio]；与CMV启动子驱动表达的相同载体相比，表达量约低6倍。与对照组EIAV载体相比，在视网膜下注射表达小鼠内皮抑素的EIAV载体，或由CMV或VMD2启动子驱动的血管抑素联合表达EIAV载体，可显著抑制Bruch膜激光诱导破裂部位的脉络膜新生血管（NV）。这些数据支持继续进行基于eiav的脉络膜NV基因治疗的临床研究，使用VMD2启动子选择性地驱动RPE细胞中内皮抑制素和血管抑制素的组合表达。
Equine infectious anemia virus (EIAV) is a nonprimate lentivirus that does not cause human disease. Subretinal injection into mice of a recombinant EIAV lentiviral vector in which lacZ is driven by a CMV promoter (EIAV CMV LacZ) resulted in rapid and strong expression of LacZ in retinal pigmented epithelial (RPE) cells and some other cells including ganglion cells, resulting in the presence of 5-bromo-4-chloro-3-indolyl-beta-d-galactopyranoside within the optic nerve. Substitution of the RPE-specific promoter from the vitelliform macular dystrophy (VMD2) gene for the CMV promoter resulted in prolonged (at least 1 year) expression of LacZ that was restricted to RPE cells, albeit reduced 6-to 10-fold compared with the CMV promoter. Similarly, the amount of FLAG-tagged endostatin detected in eyes injected with the EIAV VMD2 Endo(FLAG) vector was similar to that seen in eyes injected with a vector that expressed both endostatin and angiostatin [EIAV VMD2 Endo(FLAG)/Angio]; expression was approximately 6-fold lower than with identical vectors in which the CMV promoter drove expression. Compared with murine eyes treated with a control EIAV vector, subretinal injection of EIAV vectors expressing murine endostatin alone or in combination with angiostatin driven by either the CMV or VMD2 promoter caused significant suppression of choroidal neovascularization (NV) at laser-induced rupture sites in Bruch's membrane. These data support proceeding toward clinical studies with EIAV-based gene therapy for choroidal NV, using the VMD2 promoter to selectively drive expression of a combination of endostatin and angiostatin in RPE cells.